Natural Mycoplasma Infection Reduces Expression of Pro-Inflammatory Cytokines in Response to Ovine Footrot Pathogens.
Natural Mycoplasma Infection Reduces Expression of Pro-Inflammatory Cytokines in Response to Ovine Footrot Pathogens.
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DOI：
10.3390/ani12233235
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发表时间：
2022-11-22
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影响因子：
--
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羊蹄癣是一种痛苦的传染性疾病的蹄。由细菌Dichelobacteriumnodosus引起，有越来越多的证据表明，其他细菌在病变的起始中起着重要作用。我们之前已经确定发酵支原体与该疾病高度相关，并希望了解对与足癣相关的其他细菌的局部免疫反应的免疫抑制作用。我们从屠宰场收集的健康足部活检中建立了绵羊皮肤细胞的原代细胞培养物。初始细胞自然感染发酵支原体，分离后用抗生素处理以产生无支原体细胞系。用不同的细菌刺激不同的培养物，并在两种条件下评估mRNA和蛋白质的释放。刺激导致M.无发酵剂的细胞;然而，这不对应于蛋白质释放。自然感染M.发酵剂对刺激几乎没有反应。因此，我们得出结论，绵羊皮肤细胞感染M。发酵素对抑制对其它细菌的免疫应答的刺激的应答降低。这提供了一个重要的洞察多种病原体对宿主对腐蹄病的反应的影响。绵羊足腐病是一种复杂的多因素传染病，会导致绵羊跛行，并造成重大的福利和经济后果。节瘤叉鞭菌是主要致病菌;然而，腐蹄病是一种多微生物疾病，与坏死梭杆菌、发酵支原体和溶砷卟啉单胞菌也相关。对宿主反应的了解有限。促炎介质白细胞介素（IL）-1β和C-X-C基序趋化因子配体8（CXCL 8）已被证明在D.真皮成纤维细胞和趾间皮肤外植体模型中的结节。为了进一步了解绵羊皮肤对细菌刺激的反应，并建立对所鉴定的细胞因子和趋化因子的作用的理解，分离、培养并刺激原代绵羊趾间成纤维细胞和角质形成细胞。用LPS、D. nodosus或F. necrophorum中IL-1β和CXCL 8的转录水平增加。无发酵剂的细胞。然而，仅观察到CXCL 8蛋白释放的增加。尽管IL-1β mRNA增加，但未检测到IL-1β蛋白释放，表明当分离培养原代细胞时，可能缺乏细胞内前IL-1β加工的信号。自然感染M.发酵菌对LPS几乎没有反应，D. nodosus制剂或热灭活F.尸斑原代单细胞培养模型补充了离体器官培养模型，以研究宿主对D的反应的不同方面。结节状的感染M.发酵素对实验性细菌刺激的反应降低。然而，在腐蹄病的情况下，支原体属。与患病的脚有关，这种自然感染对多种病原体对宿主反应的影响提供了重要的见解。
Ovine footrot is a painful contagious disease of the hoof. Caused by the bacterium Dichelobacter nodosus, there is mounting evidence that other bacteria play an important role in the initiation of the lesions. We had previously established Mycoplasma fermentans as being highly associated with the disease and wanted to understand the immune-dampening effects on the local immune response to other bacteria associated with footrot. We created primary cell cultures of ovine skin cells from healthy foot biopsies collected at an abattoir. The initial cells were naturally infected with Mycoplasma fermentans, which, after isolation, were treated with antibiotics to create a Mycoplasma-free line. The different cultures were stimulated with different bacteria, and the mRNA and protein release were assessed under both conditions. The stimulation resulted in an increased expression of key immune indicators in the M. fermentans-free cells; however, this did not correspond to a protein release. The skin cells naturally infected with M. fermentans showed little response to stimulation. Therefore, we conclude Ovine skin cells infected with M. fermentans have a reduced response to stimulation dampening the immune response to other bacteria. This provides an important insight into the impact of multiple pathogens on the host response to footrot. Ovine footrot is a complex multifactorial infectious disease, causing lameness in sheep with major welfare and economic consequences. Dichelobacter nodosus is the main causative bacterium; however, footrot is a polymicrobial disease with Fusobacterium necrophorum, Mycoplasma fermentans and Porphyromonas asaccharolytica also associated. There is limited understanding of the host response involved. The proinflammatory mediators, interleukin (IL)-1β and C-X-C Motif Chemokine Ligand 8 (CXCL8), have been shown to play a role in the early response to D. nodosus in dermal fibroblasts and interdigital skin explant models. To further understand the response of ovine skin to bacterial stimulation, and to build an understanding of the role of the cytokines and chemokines identified, primary ovine interdigital fibroblasts and keratinocytes were isolated, cultured and stimulated. The expression of mRNA and protein release of CXCL8 and IL-1β were measured after stimulation with LPS, D. nodosus or F. necrophorum, which resulted in increased transcript levels of IL-1β and CXCL8 in the M. fermentans-free cells. However, only an increase in the CXCL8 protein release was observed. No IL-1β protein release was detected, despite increases in IL-1β mRNA, suggesting the signal for intracellular pre-IL-1β processing may be lacking when culturing primary cells in isolation. The keratinocytes and fibroblasts naturally infected with M. fermentans showed little response to the LPS, a range of D. nodosus preparations or heat-inactivated F. necrophorum. Primary single cell culture models complement ex vivo organ culture models to study different aspects of the host response to D. nodosus. The ovine keratinocytes and fibroblasts infected with M. fermentans had a reduced response to the experimental bacterial stimulation. However, in the case of footrot where Mycoplasma spp. are associated with diseased feet, this natural infection gives important insights into the impact of multiple pathogens on the host response.
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影响因子：
10
作者：
Nolan TJ;Gadsby NJ;Hellyer TP;Templeton KE;McMullan R;McKenna JP;Rennie J;Robb CT;Walsh TS;Rossi AG;Conway Morris A;Simpson AJ
通讯作者：
Simpson AJ
影响因子：
5.7
作者：
Maboni G;Davenport R;Sessford K;Baiker K;Jensen TK;Blanchard AM;Wattegedera S;Entrican G;Tötemeyer S
通讯作者：
Tötemeyer S
影响因子：
1.8
作者：
Davenport R;Heawood C;Sessford K;Baker M;Baiker K;Blacklaws B;Kaler J;Green L;Tötemeyer S
通讯作者：
Tötemeyer S
影响因子：
0.8
作者：
EGERTON, JR;ROBERTS, DS;PARSONSON, IM
通讯作者：
PARSONSON, IM
影响因子：
1.4
作者：
Haig, D;Deane, D;Reid, HW
通讯作者：
Reid, HW